Head-to-head comparison of BAM15, semaglutide, rosiglitazone, NEN, and calorie restriction on metabolic physiology in female db/db mice

Abstract

Metabolic disorders such as type 2 diabetes, fatty liver disease, hyperlipidemia, and obesity commonly co-occur but clinical treatment options do not effectively target all disorders. Calorie restriction, semaglutide, rosiglitazone, and mitochondrial uncouplers have all demonstrated efficacy against one or more obesity-related metabolic disorders, but it currently remains unclear which therapeutic strategy best targets the combination of hyperglycaemia, liver fat, hypertriglyceridemia, and adiposity. Herein we performed a head-to-head comparison of 5 treatment interventions in the female db/db mouse model of severe metabolic disease. Treatments included ∼60 % calorie restriction (CR), semaglutide, rosiglitazone, BAM15, and niclosamide ethanolamine (NEN). Results showed that BAM15 and CR improved body weight and liver steatosis to levels superior to semaglutide, NEN, and rosiglitazone, while BAM15, semaglutide, and rosiglitazone improved glucose tolerance better than CR and NEN. BAM15, CR, semaglutide, and rosiglitazone all had efficacy against hypertriglyceridaemia. These data provide a comprehensive head-to-head comparison of several key treatment strategies for metabolic disease and highlight the efficacy of mitochondrial uncoupling to correct multiple facets of the metabolic disease milieu in female db/db mice.

Keywords: Calorie restriction; Diabetes; GLP-1; Mitochondrial uncoupling; Obesity.

Fig. 1.

BAM15 increased oxygen consumption in female db/db mice. Oxygen consumption normalised to lean body mass (A), respiratory exchange ratio (RER, B) and locomotor activity as number of laser beam breaks (C). Pre-treatment refers to the 160 min before oral gavage and post-treatment refers to the 160 min following oral bolus dose of vehicle or BAM15. * indicates p < 0.05 as assessed by Two-Way ANOVA. Graphs show mean ± SEM. n = 6–7 per group.

Fig. 2.

Body weight and composition of female db/+ and db/db mice after 4 weeks of treatment. Body weight (A), fat mass (B), fat-free lean mass (C), percentage fat mass (D), and percentage fat-free lean mass (E). Statistical significance was analysed by One-Way ANOVA with Tukey’s multiple comparisons test, comparing each group to every other group. Only significant changes (p < 0.05) compared to db/db control are shown, indicated by *. Graphs show mean ± SEM. n = 7–9 mice per group.

Fig. 3.

Tissue wet weights in female db/+ and db/db mice after 4 weeks of treatment. Weights of single gonadal and inguinal fat pads (A) and relative fat pad weight normalised to body weight (B). Quadriceps mass as absolute weights (C) and normalised to body weight (D). Statistical significance was analysed by One-Way ANOVA with Tukey’s multiple comparisons test, comparing each group to every other group. Only significant changes (p < 0.05) compared to db/db control are shown, indicated by *. Graphs show mean ± SEM. n = 6–9 mice per group.

Fig. 4.

Glucose tolerance, blood glucose and plasma insulin levels in female db/+ and db/db mice after 4 weeks of treatment. Glucose tolerance curve (A) and area under the curve (AUC) (B). Random-fed (C) and fasting (D) blood glucose concentrations. Random-fed (E) and fasting (F) plasma insulin concentrations. Calculated HOMA-IR (G). Statistical significance was analysed by One-Way ANOVA with Tukey’s multiple comparisons test, comparing each group to every other group. Only significant changes (p < 0.05) compared to db/db control are shown, indicated by *. Graphs show mean ± SEM. n = 7–9 mice per group.

Fig. 5.

Liver and serum lipid contents in female db/+ and db/db mice after 4 weeks of treatment. Representative images of liver sections stained with haematoxylin and eosin (A). Liver wet weight in grams (B) and normalised over total body weight (C). Liver triglyceride content per gram of tissue (D) and total triglyceride in liver (E). Liver cholesterol content per gram of tissue (F) and total cholesterol in liver (G). Serum triglyceride (H) and cholesterol (I) concentrations. Statistical significance was analysed by One-Way ANOVA with Tukey’s multiple comparisons test, comparing each group to every other group. Only significant changes (p < 0.05) compared to db/db control are shown, indicated by *. Graphs show mean ± SEM. n = 7–9 mice per group.

Fig. 6.

Summary of major effects of drug treatment and calorie restriction in female db/db mice. One arrow = moderate decrease, two arrows = marked decrease, hyphen = no change.